Showing papers in "Biotechnology for Biofuels in 2021"
TL;DR: A waste biorefinery-circular bioeconomy strategy represents a low carbon economy by reducing greenhouse gases footprint, and holds great prospects for a sustainable and greener world.
Abstract: Global issues such as environmental problems and food security are currently of concern to all of us Circular bioeconomy is a promising approach towards resolving these global issues The production of bioenergy and biomaterials can sustain the energy–environment nexus as well as substitute the devoid of petroleum as the production feedstock, thereby contributing to a cleaner and low carbon environment In addition, assimilation of waste into bioprocesses for the production of useful products and metabolites lead towards a sustainable circular bioeconomy This review aims to highlight the waste biorefinery as a sustainable bio-based circular economy, and, therefore, promoting a greener environment Several case studies on the bioprocesses utilising waste for biopolymers and bio-lipids production as well as bioprocesses incorporated with wastewater treatment are well discussed The strategy of waste biorefinery integrated with circular bioeconomy in the perspectives of unravelling the global issues can help to tackle carbon management and greenhouse gas emissions A waste biorefinery–circular bioeconomy strategy represents a low carbon economy by reducing greenhouse gases footprint, and holds great prospects for a sustainable and greener world
132 citations
TL;DR: In this paper, the most recent advances in degradation and conversion of lignin to value-added bioproducts catalyzed by microbes and enzymes were summarized, and new insights for future work to overcome the heterogeneity and recalcitrance of Lignin and convert it to value added products by microbial and enzymatic catalysis were proposed.
Abstract: Lignin, the most abundant renewable aromatic compound in nature, is an excellent feedstock for value-added bioproducts manufacturing; while the intrinsic heterogeneity and recalcitrance of which hindered the efficient lignin biorefinery and utilization. Compared with chemical processing, bioprocessing with microbial and enzymatic catalysis is a clean and efficient method for lignin depolymerization and conversion. Generally, lignin bioprocessing involves lignin decomposition to lignin-based aromatics via extracellular microbial enzymes and further converted to value-added bioproducts through microbial metabolism. In the review, the most recent advances in degradation and conversion of lignin to value-added bioproducts catalyzed by microbes and enzymes were summarized. The lignin-degrading microorganisms of white-rot fungi, brown-rot fungi, soft-rot fungi, and bacteria under aerobic and anaerobic conditions were comparatively analyzed. The catalytic metabolism of the microbial lignin-degrading enzymes of laccase, lignin peroxidase, manganese peroxidase, biphenyl bond cleavage enzyme, versatile peroxidase, and β-etherize was discussed. The microbial metabolic process of H-lignin, G-lignin, S-lignin based derivatives, protocatechuic acid, and catechol was reviewed. Lignin was depolymerized to lignin-derived aromatic compounds by the secreted enzymes of fungi and bacteria, and the aromatics were converted to value-added compounds through microbial catalysis and metabolic engineering. The review also proposes new insights for future work to overcome the recalcitrance of lignin and convert it to value-added bioproducts by microbial and enzymatic catalysis.
78 citations
TL;DR: In this paper, the authors summarize the different sources of plant biomass, the evolving technologies for treating it, and the various products derived from plant biomass and the challenges inherent in the valorization of the plant biomass used in high-value-added products.
Abstract: Plant biomass is a highly abundant renewable resource that can be converted into several types of high-value-added products, including chemicals, biofuels and advanced materials. In the last few decades, an increasing number of biomass species and processing techniques have been developed to enhance the application of plant biomass followed by the industrial application of some of the products, during which varied technologies have been successfully developed. In this review, we summarize the different sources of plant biomass, the evolving technologies for treating it, and the various products derived from plant biomass. Moreover, the challenges inherent in the valorization of plant biomass used in high-value-added products are also discussed. Overall, with the increased use of plant biomass, the development of treatment technologies, and the solution of the challenges raised during plant biomass valorization, the value-added products derived from plant biomass will become greater in number and more valuable.
78 citations
TL;DR: A review of the potential utilization of engineered strains to produce biofuel and gives prospects for improvement in metabolic engineering for new strain development using advanced technologies can be found in this article, where a wide range of novel compounds have been manufactured through engineering metabolic pathways or endogenous metabolism optimizations by metabolic engineers.
Abstract: The issues of global warming, coupled with fossil fuel depletion, have undoubtedly led to renewed interest in other sources of commercial fuels. The search for renewable fuels has motivated research into the biological degradation of lignocellulosic biomass feedstock to produce biofuels such as bioethanol, biodiesel, and biohydrogen. The model strain for biofuel production needs the capability to utilize a high amount of substrate, transportation of sugar through fast and deregulated pathways, ability to tolerate inhibitory compounds and end products, and increased metabolic fluxes to produce an improved fermentation product. Engineering microbes might be a great approach to produce biofuel from lignocellulosic biomass by exploiting metabolic pathways economically. Metabolic engineering is an advanced technology for the construction of highly effective microbial cell factories and a key component for the next-generation bioeconomy. It has been extensively used to redirect the biosynthetic pathway to produce desired products in several native or engineered hosts. A wide range of novel compounds has been manufactured through engineering metabolic pathways or endogenous metabolism optimizations by metabolic engineers. This review is focused on the potential utilization of engineered strains to produce biofuel and gives prospects for improvement in metabolic engineering for new strain development using advanced technologies.
69 citations
TL;DR: In this paper, surface lignin (SL) was extracted from dilute acid-pretreated bamboo residues (DAP-BR) by various organic reagents and the residual Lignin in extracted DAP-br was obtained by the milled wood lignins (MWL) method.
Abstract: During the dilute acid pretreatment process, the resulting pseudo-lignin and lignin droplets deposited on the surface of lignocellulose and inhibit the enzymatic digestibility of cellulose in lignocellulose. However, how these lignins interact with cellulase enzymes and then affect enzymatic hydrolysis is still unknown. In this work, different fractions of surface lignin (SL) obtained from dilute acid-pretreated bamboo residues (DAP-BR) were extracted by various organic reagents and the residual lignin in extracted DAP-BR was obtained by the milled wood lignin (MWL) method. All of the lignin fractions obtained from DAP-BR were used to investigate the mechanism for interaction between lignin and cellulase using surface plasmon resonance (SPR) technology to understand how they affect enzymatic hydrolysis The results showed that removing surface lignin significantly decreased the yield for enzymatic hydrolysis DAP-BR from 36.5% to 18.6%. The addition of MWL samples to Avicel inhibited its enzymatic hydrolysis, while different SL samples showed slight increases in enzymatic digestibility. Due to the higher molecular weight and hydrophobicity of MWL samples versus SL samples, a stronger affinity for MWL (KD = 6.8–24.7 nM) was found versus that of SL (KD = 39.4–52.6 nM) by SPR analysis. The affinity constants of all tested lignins exhibited good correlations (r > 0.6) with the effects on enzymatic digestibility of extracted DAP-BR and Avicel. This work revealed that the surface lignin on DAP-BR is necessary for maintaining enzyme digestibility levels, and its removal has a negative impact on substrate digestibility.
63 citations
TL;DR: In this paper, the effect of residual lignin on enzymatic hydrolysis of lignocellulose feedstocks is discussed. But the authors focus on exploring the interaction between Lignin-enzyme interactions to mitigate the negative impact of LIGNIN and reduce the cost of enzymatically hydrolyzing lignosulfonate.
Abstract: Enzymatic hydrolysis of lignocellulose for bioethanol production shows a great potential to remit the rapid consumption of fossil fuels, given the fact that lignocellulose feedstocks are abundant, cost-efficient, and renewable. Lignin results in low enzymatic saccharification by forming the steric hindrance, non-productive adsorption of cellulase onto lignin, and deactivating the cellulase. In general, the non-productive binding of cellulase on lignin is widely known as the major cause for inhibiting the enzymatic hydrolysis. Pretreatment is an effective way to remove lignin and improve the enzymatic digestibility of lignocellulose. Along with removing lignin, the pretreatment can modify the lignin structure, which significantly affects the non-productive adsorption of cellulase onto lignin. To relieve the inhibitory effect of lignin on enzymatic hydrolysis, enormous efforts have been made to elucidate the correlation of lignin structure with lignin-enzyme interactions but with different views. In addition, contrary to the traditional belief that lignin inhibits enzymatic hydrolysis, in recent years, the addition of water-soluble lignin such as lignosulfonate or low molecular-weight lignin exerts a positive effect on enzymatic hydrolysis, which gives a new insight into the lignin-enzyme interactions. For throwing light on their structure-interaction relationship during enzymatic hydrolysis, the effect of residual lignin in substrate and introduced lignin in hydrolysate on enzymatic hydrolysis are critically reviewed, aiming at realizing the targeted regulation of lignin structure for improving the saccharification of lignocellulose. The review is also focused on exploring the lignin-enzyme interactions to mitigate the negative impact of lignin and reducing the cost of enzymatic hydrolysis of lignocellulose.
55 citations
TL;DR: In this paper, a yeast consortium, NYC-1, which was constructed included the manganese-dependent peroxidase producing oleaginous strains Meyerozyma caribbica, Meyerzyma guilliermondii, Debaryomyces hansenii, and Vanrija humicola, and showed efficient azo dyes decolorization.
Abstract: Textile industry represents one prevalent activity worldwide, generating large amounts of highly contaminated and rich in azo dyes wastewater, with severe effects on natural ecosystems and public health. However, an effective and environmentally friendly treatment method has not yet been implemented, while concurrently, the increasing demand of modern societies for adequate and sustainable energy supply still remains a global challenge. Under this scope, the purpose of the present study was to isolate promising species of yeasts inhabiting wood-feeding termite guts, for combined azo dyes and textile wastewater bioremediation, along with biodiesel production. Thirty-eight yeast strains were isolated, molecularly identified and subsequently tested for desired enzymatic activity, lipid accumulation, and tolerance to lignin-derived metabolites. The most promising species were then used for construction of a novel yeast consortium, which was further evaluated for azo dyes degradation, under various culture conditions, dye levels, as well as upon the addition of heavy metals, different carbon and nitrogen sources, and lastly agro-waste as an inexpensive and environmentally friendly substrate alternative. The novel yeast consortium, NYC-1, which was constructed included the manganese-dependent peroxidase producing oleaginous strains Meyerozyma caribbica, Meyerozyma guilliermondii, Debaryomyces hansenii, and Vanrija humicola, and showed efficient azo dyes decolorization, which was further enhanced depending on the incubation conditions. Furthermore, enzymatic activity, fatty acid profile and biodiesel properties were thoroughly investigated. Lastly, a dye degradation pathway coupled to biodiesel production was proposed, including the formation of phenol-based products, instead of toxic aromatic amines. In total, this study might be the first to explore the application of MnP and lipid-accumulating yeasts for coupling dye degradation and biodiesel production.
52 citations
TL;DR: A novel two-domain cellulose-active family AA10 LPMO from a marine actinomycete is described and the observed differences between ascorbic acid and gallic acid show a way of making L PMO reactions less copper-dependent and illustrate that reductant effects on LPMo action need to be interpreted with great caution.
Abstract: Lytic polysaccharide monooxygenases (LPMOs) are monocopper enzymes that catalyze oxidative depolymerization of industrially relevant crystalline polysaccharides, such as cellulose, in a reaction that depends on an electron donor and O2 or H2O2. While it is well known that LPMOs can utilize a wide variety of electron donors, the variation in reported efficiencies of various LPMO-reductant combinations remains largely unexplained. In this study, we describe a novel two-domain cellulose-active family AA10 LPMO from a marine actinomycete, which we have used to look more closely at the effects of the reductant and copper ions on the LPMO reaction. Our results show that ascorbate-driven LPMO reactions are extremely sensitive to very low amounts (micromolar concentrations) of free copper because reduction of free Cu(II) ions by ascorbic acid leads to formation of H2O2, which speeds up the LPMO reaction. In contrast, the use of gallic acid yields steady reactions that are almost insensitive to the presence of free copper ions. Various experiments, including dose–response studies with the enzyme, showed that under typically used reaction conditions, the rate of the reaction is limited by LPMO-independent formation of H2O2 resulting from oxidation of the reductant. The strong impact of low amounts of free copper on LPMO reactions with ascorbic acid and O2, i.e. the most commonly used conditions when assessing LPMO activity, likely explains reported variations in LPMO rates. The observed differences between ascorbic acid and gallic acid show a way of making LPMO reactions less copper-dependent and illustrate that reductant effects on LPMO action need to be interpreted with great caution. In clean reactions, with minimized generation of H2O2, the (O2-driven) LPMO reaction is exceedingly slow, compared to the much faster peroxygenase reaction that occurs when adding H2O2.
51 citations
TL;DR: A review of state-of-the-art research in the field of process systems engineering with a particular focus on biodiesel production including process design and simulation, sustainability evaluation, optimization and supply chain management is presented in this paper.
Abstract: The overwhelming concerns due to over exploitation of fossil resources necessitate the utilization of alternative energy resources. Biodiesel has been considered as one of the most adaptable alternative to fossil-derived diesel with similar properties and numerous environmental benefits. Although there are various approaches for biodiesel production, development of cost-effective and robust catalyst with efficient production methods and utilization of a variety of feedstock could be the optimum solution to bring down the production cost. Considering the complexity of biodiesel production processes, process design, quantitative evaluation and optimization of the biodiesel from whole systems perspectives is essential for unlocking the complexity and enhancing the system performances. Process systems engineering offers an efficient approach to design and optimize biodiesel manufacturing systems by using a variety of tools. This review reflects state-of-the-art biodiesel research in the field of process systems engineering with a particular focus on biodiesel production including process design and simulation, sustainability evaluation, optimization and supply chain management. This review also highlights the challenges and opportunities for the development of potentially sustainable and eco-friendly enzymatic biodiesel technology.
46 citations
TL;DR: In this article, the authors discussed different pretreatment technologies of hydrolysis and their restrictions and showed that different pretreatments have varying effects on lignin, cellulose, and hemicellulose degradation and biogas yield of different substrate.
Abstract: Population increase and industrialization has resulted in high energy demand and consumptions, and presently, fossil fuels are the major source of staple energy, supplying 80% of the entire consumption. This has contributed immensely to the greenhouse gas emission and leading to global warming, and as a result of this, there is a tremendous urgency to investigate and improve fresh and renewable energy sources worldwide. One of such renewable energy sources is biogas that is generated by anaerobic fermentation that uses different wastes such as agricultural residues, animal manure, and other organic wastes. During anaerobic digestion, hydrolysis of substrates is regarded as the most crucial stage in the process of biogas generation. However, this process is not always efficient because of the domineering stableness of substrates to enzymatic or bacteria assaults, but substrates' pretreatment before biogas production will enhance biogas production. The principal objective of pretreatments is to ease the accessibility of the enzymes to the lignin, cellulose, and hemicellulose which leads to degradation of the substrates. Hence, the use of pretreatment for catalysis of lignocellulose substrates is beneficial for the production of cost-efficient and eco-friendly process. In this review, we discussed different pretreatment technologies of hydrolysis and their restrictions. The review has shown that different pretreatments have varying effects on lignin, cellulose, and hemicellulose degradation and biogas yield of different substrate and the choice of pretreatment technique will devolve on the intending final products of the process.
45 citations
TL;DR: In this article, the barriers and solutions to rapid development of genetic transformation tools in new hosts, with a major focus on Restriction-Modification systems, are reviewed, and tools and approaches used for efficient gene deletion, DNA insertion, and heterologous gene expression.
Abstract: Non-model microorganisms often possess complex phenotypes that could be important for the future of biofuel and chemical production. They have received significant interest the last several years, but advancement is still slow due to the lack of a robust genetic toolbox in most organisms. Typically, “domestication” of a new non-model microorganism has been done on an ad hoc basis, and historically, it can take years to develop transformation and basic genetic tools. Here, we review the barriers and solutions to rapid development of genetic transformation tools in new hosts, with a major focus on Restriction-Modification systems, which are a well-known and significant barrier to efficient transformation. We further explore the tools and approaches used for efficient gene deletion, DNA insertion, and heterologous gene expression. Finally, more advanced and high-throughput tools are now being developed in diverse non-model microbes, paving the way for rapid and multiplexed genome engineering for biotechnology.
TL;DR: In this article, the authors focus on the challenges associated with sustainable production of lignocellulosic nanoscale polymeric materials (NPMs) need to be addressed.
Abstract: Plant-biomass-based nanomaterials have attracted great interest recently for their potential to replace petroleum-sourced polymeric materials for sustained economic development. However, challenges associated with sustainable production of lignocellulosic nanoscale polymeric materials (NPMs) need to be addressed. Producing materials from lignocellulosic biomass is a value-added proposition compared with fuel-centric approach. This report focuses on recent progress made in understanding NPMs—specifically lignin nanoparticles (LNPs) and cellulosic nanomaterials (CNMs)—and their sustainable production. Special attention is focused on understanding key issues in nano-level deconstruction of cell walls and utilization of key properties of the resultant NPMs to allow flexibility in production to promote sustainability. Specifically, suitable processes for producing LNPs and their potential for scaled-up production, along with the resultant LNP properties and prospective applications, are discussed. In the case of CNMs, terminologies such as cellulose nanocrystals (CNCs) and cellulose nanofibrils (CNFs) used in the literature are examined. The term cellulose nano-whiskers (CNWs) is used here to describe a class of CNMs that has a morphology similar to CNCs but without specifying its crystallinity, because most applications of CNCs do not need its crystalline characteristic. Additionally, progress in enzymatic processing and drying of NPMs is also summarized. Finally, the report provides some perspective of future research that is likely to result in commercialization of plant-based NPMs.
TL;DR: In this paper, an overview of ethanol separation via pervaporation primarily concentrates on transport mechanisms, fabrication methods, and membrane materials; the research and development of polymeric, inorganic and mixed matrix membranes are reviewed from the perspective of membrane materials as well as modification methods.
Abstract: Bioethanol as a renewable energy resource plays an important role in alleviating energy crisis and environmental protection. Pervaporation has achieved increasing attention because of its potential to be a useful way to separate ethanol from the biomass fermentation process. This overview of ethanol separation via pervaporation primarily concentrates on transport mechanisms, fabrication methods, and membrane materials. The research and development of polymeric, inorganic, and mixed matrix membranes are reviewed from the perspective of membrane materials as well as modification methods. The recovery performance of the existing pervaporation membranes for ethanol solutions is compared, and the approaches to further improve the pervaporation performance are also discussed. Overall, exploring the possibility and limitation of the separation performance of PV membranes for ethanol extraction is a long-standing topic. Collectively, the quest is to break the trade-off between membrane permeability and selectivity. Based on the facilitated transport mechanism, further exploration of ethanol-selective membranes may focus on constructing a well-designed microstructure, providing active sites for facilitating the fast transport of ethanol molecules, hence achieving both high selectivity and permeability simultaneously. Finally, it is expected that more and more successful research could be realized into commercial products and this separation process will be deployed in industrial practices in the near future.
TL;DR: In this paper, furoic acid was employed to pre-process sugarcane bagasse to recover the xylooligosaccharides fraction from hemicellulose and boost the subsequent cellulose saccharification.
Abstract: Pretreatment is the key step for utilizing lignocellulosic biomass, which can extract cellulose from lignin and disrupt its recalcitrant crystalline structure to allow much more effective enzymatic hydrolysis; and organic acids pretreatment with dual benefic for generating xylooligosaccharides and boosting enzymatic hydrolysis has been widely used in adding values to lignocellulose materials. In this work, furoic acid, a novel recyclable organic acid as catalyst, was employed to pretreat sugarcane bagasse to recover the xylooligosaccharides fraction from hemicellulose and boost the subsequent cellulose saccharification. The FA-assisted hydrolysis of sugarcane bagasse using 3% furoic acid at 170 °C for 15 min resulted in the highest xylooligosaccharides yield of 45.6%; subsequently, 83.1 g/L of glucose was harvested by a fed-batch operation with a solid loading of 15%. Overall, a total of 120 g of xylooligosaccharides and 335 g glucose could be collected from 1000 g sugarcane bagasse starting from the furoic acid pretreatment. Furthermore, furoic acid can be easily recovered by cooling crystallization. This work put forward a novel furoic acid pretreatment method to convert sugarcane bagasse into xylooligosaccharides and glucose, which provides a strategy that the sugar and nutraceutical industries can be used to reduce the production cost. The developed process showed that the yields of xylooligosaccharides and byproducts were controllable by shortening the reaction time; meanwhile, the recyclability of furoic acid also can potentially reduce the pretreatment cost and potentially replace the traditional mineral acids pretreatment.
TL;DR: In this paper, the effect of individual inhibitors in inhibitor-rich lignocellulose hydrolysates has been investigated by spiking higher concentrations of each compound in a concentration range relevant for industrial enzymes.
Abstract: Presence of inhibitory chemicals in lignocellulose hydrolysates is a major hurdle for production of second-generation bioethanol. Especially cheaper pre-treatment methods that ensure an economical viable production process generate high levels of these inhibitory chemicals. The effect of several of these inhibitors has been extensively studied with non-xylose-fermenting laboratory strains, in synthetic media, and usually as single inhibitors, or with inhibitor concentrations much higher than those found in lignocellulose hydrolysates. However, the relevance of individual inhibitors in inhibitor-rich lignocellulose hydrolysates has remained unclear. The relative importance for inhibition of ethanol fermentation by two industrial second-generation yeast strains in five lignocellulose hydrolysates, from bagasse, corn cobs and spruce, has now been investigated by spiking higher concentrations of each compound in a concentration range relevant for industrial hydrolysates. The strongest inhibition was observed with industrially relevant concentrations of furfural causing partial inhibition of both D-glucose and D-xylose consumption. Addition of 3 or 6 g/L furfural strongly reduced the ethanol titer obtained with strain MD4 in all hydrolysates evaluated, in a range of 34 to 51% and of 77 to 86%, respectively. This was followed by 5-hydroxymethylfurfural, acetic acid and formic acid, for which in general, industrially relevant concentrations caused partial inhibition of D-xylose fermentation. On the other hand, spiking with levulinic acid, 4-hydroxybenzaldehyde, 4-hydroxybenzoic acid or vanillin caused little inhibition compared to unspiked hydrolysate. The further evolved MD4 strain generally showed superior performance compared to the previously developed strain GSE16-T18. The results highlight the importance of individual inhibitor evaluation in a medium containing a genuine mix of inhibitors as well as the ethanol that is produced by the fermentation. They also highlight the potential of increasing yeast inhibitor tolerance for improving industrial process economics.
TL;DR: A comprehensive overview of fluorescence label-based imaging techniques and a variety of probing methods for plant cell wall analysis can be found in this article, where the authors elaborate on what are the most important considerations when applying a particular technique for plants, the potential for future development, and how the plantcell wall field might be inspired by advances in the biomedical and general cell biology fields.
Abstract: Plant cell wall-derived biomass serves as a renewable source of energy and materials with increasing importance. The cell walls are biomacromolecular assemblies defined by a fine arrangement of different classes of polysaccharides, proteoglycans, and aromatic polymers and are one of the most complex structures in Nature. One of the most challenging tasks of cell biology and biomass biotechnology research is to image the structure and organization of this complex matrix, as well as to visualize the compartmentalized, multiplayer biosynthetic machineries that build the elaborate cell wall architecture. Better knowledge of the plant cells, cell walls, and whole tissue is essential for bioengineering efforts and for designing efficient strategies of industrial deconstruction of the cell wall-derived biomass and its saccharification. Cell wall-directed molecular probes and analysis by light microscopy, which is capable of imaging with a high level of specificity, little sample processing, and often in real time, are important tools to understand cell wall assemblies. This review provides a comprehensive overview about the possibilities for fluorescence label-based imaging techniques and a variety of probing methods, discussing both well-established and emerging tools. Examples of applications of these tools are provided. We also list and discuss the advantages and limitations of the methods. Specifically, we elaborate on what are the most important considerations when applying a particular technique for plants, the potential for future development, and how the plant cell wall field might be inspired by advances in the biomedical and general cell biology fields.
TL;DR: In this article, the authors provided an in-depth assessment of Ethiopia's biomass energy availability, potential, challenges, and prospects, and the most effective techniques for producing and utilizing alternate energy sources were also explored.
Abstract: Despite enormous challenges in accessing sustainable energy supplies and advanced energy technologies, Ethiopia has one of the world's fastest growing economies. The development of renewable energy technology and the building of a green legacy in the country are being prioritized. The total installed capacity for electricity generation in Ethiopia is 4324.3 MW as on October, 2018. Renewable energy accounts for 96.5% of total generation; however, despite the county's enormous biomass energy potential, only 0.58% of power is generated using biomass. Ethiopia has surplus woody biomass, crop residue and animal dung resources which comprise about 141.8 million metric tons of biomass availability per year. At present the exploited potential is about 71.9 million metric tons per year. This review paper provides an in-depth assessment of Ethiopia's biomass energy availability, potential, challenges, and prospects. The findings show that, despite Ethiopia's vast biomass resource potential, the current use of modern energy from biomass is still limited. As a result, this study supports the use of biomass-based alternative energy sources without having a negative impact on the socioeconomic system or jeopardizing food security or the environment. This finding also shows the challenges, opportunities and possible solutions to tackle the problem to expand alternative energy sources. The most effective techniques for producing and utilizing alternate energy sources were also explored. Moreover, some perspectives are given based on the challenges of using efficient energy production and sustainable uses of biomass energy in Ethiopia as it could be also implemented in other developing countries. We believe that the information in this review will shed light on the current and future prospects of biomass energy deployment in Ethiopia.
TL;DR: In this paper, the degradation profiles of the fractions were compared considering physical changes in addition to chemical conversion, and the lowest and highest saccharification yields were measured in the coarse and fine fractions, respectively, and these yields paralleled the reduction in the size and number of particles.
Abstract: Background : The recalcitrance of lignocellulosics to enzymatic saccharification has been related to many factors, including the tissue and molecular heterogeneity of the plant particles. The role of tissue heterogeneity generally assessed from plant sections is not easy to study on a large scale. In the present work, dry fractionation of ground maize shoot was performed to obtain particle fractions enriched in a specific tissue. The degradation profiles of the fractions were compared considering physical changes in addition to chemical conversion.
Results : Coarse, medium and fine fractions were produced using a dry process followed by an electrostatic separation. The physical and chemical characteristics of the fractions varied, suggesting enrichment in tissue from leaves, pith or rind. The fractions were subjected to enzymatic hydrolysis in a torus reactor designed for real-time monitoring of the number and size of the particles. Saccharification efficiency was monitored by analyzing the sugar release at different times. The lowest and highest saccharification yields were measured in the coarse and fine fractions, respectively, and these yields paralleled the reduction in the size and number of particles. The behavior of the positively- and negatively-charged particles of medium-size fractions was contrasted. Although the amount of sugar release was similar, the changes in particle size and number differed during enzymatic degradation. The reduction in the number of particles proceeded faster than that of particle size, suggesting that degradable particles were degraded to the point of disappearance with no significant erosion or fragmentation. Considering all fractions, the saccharification yield was positively correlated with the amount of water associated with [5–15 nm] pore size range at 67% moisture content while the reduction in the number of particles was inversely correlated with the amount of lignin.
Conclusion : Real-time monitoring of sugar release and changes in the number and size of the particles clearly evidenced different degradation patterns for fractions of maize shoot that could be related to tissue heterogeneity in the plant. The biorefinery process could benefit from the addition of a sorting stage to optimise the flow of biomass materials and take better advantage of the heterogeneity of the biomass.
TL;DR: In this paper, the ability of oleaginous yeast to convert crude glycerol (CG) and hemicellulose hydrolysate (HH) to microbial lipids was investigated.
Abstract: Crude glycerol (CG) and hemicellulose hydrolysate (HH) are low—value side-products of biodiesel transesterification and pulp—and paper industry or lignocellulosic ethanol production, respectively, which can be converted to microbial lipids by oleaginous yeasts. This study aimed to test the ability of oleaginous yeasts to utilise CG and HH and mixtures of them as carbon source. Eleven out of 27 tested strains of oleaginous yeast species were able to grow in plate tests on CG as sole carbon source. Among them, only one ascomycetous strain, belonging to Lipomyces starkeyi, was identified, the other 10 strains were Rhodotorula spec. When yeasts were cultivated in mixed CG/ HH medium, we observed an activation of glycerol conversion in the Rhodotorula strains, but not in L. starkeyi. Two strains—Rhodotorula toruloides CBS 14 and Rhodotorula glutinis CBS 3044 were further tested in controlled fermentations in bioreactors in different mixtures of CG and HH. The highest measured average biomass and lipid concentration were achieved with R. toruloides in 10% HH medium mixed with 55 g/L CG—19.4 g/L and 10.6 g/L, respectively, with a lipid yield of 0.25 g lipids per consumed g of carbon source. Fatty acid composition was similar to other R. toruloides strains and comparable to that of vegetable oils. There were big strain differences in the ability to convert CG to lipids, as only few of the tested strains were able to grow. Lipid production rates and yields showed that mixing GC and HH have a stimulating effect on lipid accumulation in R. toruloides and R. glutinis resulting in shortened fermentation time to reach maximum lipid concentration, which provides a new perspective on converting these low-value compounds to microbial lipids.
TL;DR: A novel MFC-assisted system was established to improve glycerol utilization by A. succinogenes for succinate and electricity production, making this system as a platform for chemicals production and electrical supply simultaneously.
Abstract: The global production of glycerol is increasing year by year since the demands of biodiesel is rising. It is benefit for high-yield succinate synthesis due to its high reducing property. A. succinogenes, a succinate-producing candidate, cannot grow on glycerol anaerobically, as it needs a terminal electron acceptor to maintain the balance of intracellular NADH and NAD+. Microbial fuel cell (MFC) has been widely used to release extra intracellular electrons. However, A. succinogenes is a non-electroactive strain which need the support of electron shuttle in MFC, and pervious research showed that acid-tolerant A. succinogenes has higher content of unsaturated fatty acids, which may be beneficial for the transmembrane transport of lipophilic electron shuttle. MFC-assisted succinate production was evaluated using neutral red as an electron shuttle to recover the glycerol utilization. First, an acid-tolerant mutant JF1315 was selected by atmospheric and room temperature plasma (ARTP) mutagenesis aiming to improve transmembrane transport of neutral red (NR). Additionally, MFC was established to increase the ratio of oxidized NR to reduced NR. By combining these two strategies, ability of JF1315 for glycerol utilization was significantly enhanced, and 23.92 g/L succinate was accumulated with a yield of 0.88 g/g from around 30 g/L initial glycerol, along with an output voltage above 300 mV. A novel MFC-assisted system was established to improve glycerol utilization by A. succinogenes for succinate and electricity production, making this system as a platform for chemicals production and electrical supply simultaneously.
TL;DR: The addition of FAST as an oxygen-independent fluorescent reporter protein expands the genetic toolbox of E. limosum and shows that FAST-tagged fusion proteins can be constructed without negatively impacting the stability, functionality, and productivity of the resulting enzyme.
Abstract: The interest in using methanol as a substrate to cultivate acetogens increased in recent years since it can be sustainably produced from syngas and has the additional benefit of reducing greenhouse gas emissions Eubacterium limosum is one of the few acetogens that can utilize methanol, is genetically accessible and, therefore, a promising candidate for the recombinant production of biocommodities from this C1 carbon source Although several genetic tools are already available for certain acetogens including E limosum, the use of brightly fluorescent reporter proteins is still limited In this study, we expanded the genetic toolbox of E limosum by implementing the fluorescence-activating and absorption shifting tag (FAST) as a fluorescent reporter protein Recombinant E limosum strains that expressed the gene encoding FAST in an inducible and constitutive manner were constructed Cultivation of these recombinant strains resulted in brightly fluorescent cells even under anaerobic conditions Moreover, we produced the biocommodities butanol and acetone from methanol with recombinant E limosum strains Therefore, we used E limosum cultures that produced FAST-tagged fusion proteins of the bifunctional acetaldehyde/alcohol dehydrogenase or the acetoacetate decarboxylase, respectively, and determined the fluorescence intensity and product concentrations during growth The addition of FAST as an oxygen-independent fluorescent reporter protein expands the genetic toolbox of E limosum Moreover, our results show that FAST-tagged fusion proteins can be constructed without negatively impacting the stability, functionality, and productivity of the resulting enzyme Finally, butanol and acetone can be produced from methanol using recombinant E limosum strains expressing genes encoding fluorescent FAST-tagged fusion proteins
TL;DR: A considerable escalation on the enzymatic activities obtained in a short period from the cultivation of the L. betulina or T. versicolor due to the enhanced microbial synergistic effects, providing a potential bioconversion route for the applications of lignin utilization.
Abstract: As one of the major components of lignocellulosic biomass, lignin has been considered as the most abundant renewable aromatic feedstock in the world. Comparing with thermal or catalytic strategies for lignin degradation, biological conversion is a promising approach featuring with mild conditions and diversity, and has received great attention nowadays. In this study, a consortium of white rot fungi composed of Lenzites betulina and Trametes versicolor was employed to enhance the ligninolytic enzyme activity of laccase (Lac) and manganese peroxidase (MnP) under microbial synergism. The maximum enzymatic activity of Lac and MnP was individually 18.06 U mL−1 and 13.58 U mL−1 along with a lignin degradation rate of 50% (wt/wt), which were achieved from batch cultivation of the consortium. The activities of Lac and MnP obtained from the consortium were both improved more than 40%, as compared with monocultures of L. betulina or T. versicolor under the same culture condition. The enhanced biodegradation performance was in accordance with the results observed from scanning electron microscope (SEM) of lignin samples before and after biodegradation, and secondary-ion mass spectrometry (SIMS). Finally, the analysis of heteronuclear single quantum coherence (HSQC) NMR and gas chromatography–mass spectrometry (GC–MS) provided a comprehensive product mapping of the lignin biodegradation, suggesting that the lignin has undergone depolymerization of the macromolecules, side-chain cleavage, and aromatic ring-opening reactions. Our results revealed a considerable escalation on the enzymatic activity obtained in a short period from the cultivation of the L. betulina or T. versicolor due to the enhanced microbial synergistic effects, providing a potential bioconversion route for lignin utilization.
TL;DR: In this article, a mixotrophic metabolic mechanism was identified in a mixture of photoautotrophic and heterotrophy in microalgal species, and the authors demonstrated that the intermediates of glycolysis could directly enter the chloroplast and replace RuBisCO-fixed CO2 to provide carbon sources for chloropplastic organic carbon metabolism under mixotroglobal growth.
Abstract: Mixotrophy can confer a higher growth rate than the sum of photoautotrophy and heterotrophy in many microalgal species Thus, it has been applied to biodiesel production and wastewater utilization However, its carbon and energy metabolic mechanism is currently poorly understood To elucidate underlying carbon and energy metabolic mechanism of mixotrophy, Chromochloris zofingiensis was employed in the present study Photosynthesis and glucose metabolism were found to operate in a dynamic balance during mixotrophic cultivation, the enhancement of one led to the lowering of the other Furthermore, compared with photoautotrophy, non-photochemical quenching and photorespiration, considered by many as energy dissipation processes, were significantly reduced under mixotrophy Comparative transcriptome analysis suggested that the intermediates of glycolysis could directly enter the chloroplast and replace RuBisCO-fixed CO2 to provide carbon sources for chloroplast organic carbon metabolism under mixotrophy Therefore, the photosynthesis rate-limiting enzyme, RuBisCO, was skipped, allowing for more efficient utilization of photoreaction-derived energy Besides, compared with heterotrophy, photoreaction-derived ATP reduced the need for TCA-derived ATP, so the glucose decomposition was reduced, which led to higher biomass yield on glucose Based on these results, a mixotrophic metabolic mechanism was identified Our results demonstrate that the intermediates of glycolysis could directly enter the chloroplast and replace RuBisCO-fixed CO2 to provide carbon for photosynthesis in mixotrophy Therefore, the photosynthesis rate-limiting enzyme, RuBisCO, was skipped in mixotrophy, which could reduce energy waste of photosynthesis while promote cell growth This finding provides a foundation for future studies on mixotrophic biomass production and photosynthetic metabolism
TL;DR: In this paper, the role of secretomics in identifying enzymes involved in lignocelluloses deconstruction, the development of enzyme cocktails and the construction of synthetic microbial consortia for biomass valorization, providing their perspectives to address the current challenges.
Abstract: The recalcitrance of lignocellulosic biomass is a major constraint to its high-value use at industrial scale. In nature, microbes play a crucial role in biomass degradation, nutrient recycling and ecosystem functioning. Therefore, the use of microbes is an attractive way to transform biomass to produce clean energy and high-value compounds. The microbial degradation of lignocelluloses is a complex process which is dependent upon multiple secreted enzymes and their synergistic activities. The availability of the cutting edge proteomics and highly sensitive mass spectrometry tools make possible for researchers to probe the secretome of microbes and microbial consortia grown on different lignocelluloses for the identification of hydrolytic enzymes of industrial interest and their substrate-dependent expression. This review summarizes the role of secretomics in identifying enzymes involved in lignocelluloses deconstruction, the development of enzyme cocktails and the construction of synthetic microbial consortia for biomass valorization, providing our perspectives to address the current challenges.
TL;DR: The E. coli AppA phytase, ApV1, contains an extra non-consecutive disulfide bond as discussed by the authors, which may result in folding complexity and decreased production in microbial systems.
Abstract: Phytases are widely used commercially as dietary supplements for swine and poultry to increase the digestibility of phytic acid. Enzyme development has focused on increasing thermostability to withstand the high temperatures during industrial steam pelleting. Increasing thermostability often reduces activity at gut temperatures and there remains a demand for improved phyases for a growing market. In this work, we present a thermostable variant of the E. coli AppA phytase, ApV1, that contains an extra non-consecutive disulfide bond. Detailed biochemical characterisation of ApV1 showed similar activity to the wild type, with no statistical differences in kcat and KM for phytic acid or in the pH and temperature activity optima. Yet, it retained approximately 50% activity after incubations for 20 min at 65, 75 and 85 °C compared to almost full inactivation of the wild-type enzyme. Production of ApV1 in Pichia pastoris (Komagataella phaffi) was much lower than the wild-type enzyme due to the presence of the extra non-consecutive disulfide bond. Production bottlenecks were explored using bidirectional promoters for co-expression of folding chaperones. Co-expression of protein disulfide bond isomerase (Pdi) increased production of ApV1 by ~ 12-fold compared to expression without this folding catalyst and restored yields to similar levels seen with the wild-type enzyme. Overall, the results show that protein engineering for enhanced enzymatic properties like thermostability may result in folding complexity and decreased production in microbial systems. Hence parallel development of improved production strains is imperative to achieve the desirable levels of recombinant protein for industrial processes.
TL;DR: In this article, the authors summarized recent progress on the production of triacylglycerol and astaxanthin by C. zofingiensis and proposed strategies for trait improvements.
Abstract: The algal lipids-based biodiesel, albeit having advantages over plant oils, still remains high in the production cost. Co-production of value-added products with lipids has the potential to add benefits and is thus believed to be a promising strategy to improve the production economics of algal biodiesel. Chromochloris zofingiensis, a unicellular green alga, has been considered as a promising feedstock for biodiesel production because of its robust growth and ability of accumulating high levels of triacylglycerol under multiple trophic conditions. This alga is also able to synthesize high-value keto-carotenoids and has been cited as a candidate producer of astaxanthin, the strongest antioxidant found in nature. The concurrent accumulation of triacylglycerol and astaxanthin enables C. zofingiensis an ideal cell factory for integrated production of the two compounds and has potential to improve algae-based production economics. Furthermore, with the advent of chromosome-level whole genome sequence and genetic tools, C. zofingiensis becomes an emerging model for studying lipid metabolism and carotenogenesis. In this review, we summarize recent progress on the production of triacylglycerol and astaxanthin by C. zofingiensis. We also update our understanding in the distinctive molecular mechanisms underlying lipid metabolism and carotenogenesis, with an emphasis on triacylglycerol and astaxanthin biosynthesis and crosstalk between the two pathways. Furthermore, strategies for trait improvements are discussed regarding triacylglycerol and astaxanthin synthesis in C. zofingiensis.
TL;DR: In this paper, the surface barrier effect of lignin on cellulose hydrolysis was isolated from unproductive binding effect of Lignin, and the analyses of surface chemistry, surface morphology and surface area were carried out to reveal the effect of various additives.
Abstract: Ethanol organosolv (EOS) pretreatment is one of the most efficient methods for boosting biomass saccharification as it can achieve an efficient fractionation of three major constituents in lignocellulose. However, lignin repolymerization often occurs in acid EOS pretreatment, which impairs subsequent enzymatic hydrolysis. This study investigated acid EOS pretreatment assisted by carbocation scavenger (2-naphthol, 2-naphthol-7-sulfonate, mannitol and syringic acid) to improve biomass fractionation, coproduction of fermentable sugars and lignin adsorbents. In addition, surface barrier effect of lignin on cellulose hydrolysis was isolated from unproductive binding effect of lignin, and the analyses of surface chemistry, surface morphology and surface area were carried out to reveal the lignin inhibition mitigating effect of various additives. Four different additives all helped mitigate lignin inhibition on cellulose hydrolysis in particular diminishing surface barrier effect, among which 2-naphthol-7-sulfonate showed the best performance in improving pretreatment efficacy, while mannitol and syringic acid could serve as novel green additives. Through the addition of 2-naphthol-7-sulfonate, selective lignin removal was increased up to 76%, while cellulose hydrolysis yield was improved by 85%. As a result, 35.78 kg cellulose and 16.63 kg hemicellulose from 100 kg poplar could be released and recovered as fermentable sugars, corresponding to a sugar yield of 78%. Moreover, 22.56 kg ethanol organosolv lignin and 17.53 kg enzymatic hydrolysis residue could be recovered as lignin adsorbents for textile dye removal, with the adsorption capacities of 45.87 and 103.09 mg g−1, respectively. Results in this work indicated proper additives could give rise to the form of less repolymerized surface lignin, which would decrease the unproductive binding of cellulase enzymes to surface lignin. Besides, the supplementation of additives (NS, MT and SA) resulted in a simultaneously increased surface area and decreased lignin coverage. All these factors contributed to the diminished surface barrier effect of lignin, thereby improving the ease of enzymatic hydrolysis of cellulose. The biorefinery process based on acidic EOS pretreatment assisted by carbocation scavenger was proved to enable the coproduction of fermentable sugars and lignin adsorbents, allowing the holistic utilization of lignocellulosic biomass for a sustainable biorefinery.
TL;DR: In this paper, a series of 2D 13C-13C correlation spectra were successfully collected using the unlabeled stems of wild-type Oryza sativa (rice), and the atomic resolution allowed to observe a large number of intramolecular cross peaks for fully revealing the polymorphic structure of cellulose and xylan.
Abstract: Multidimensional solid-state nuclear magnetic resonance (ssNMR) spectroscopy has emerged as an indispensable technique for resolving polymer structure and intermolecular packing in primary and secondary plant cell walls. Isotope (13C) enrichment provides feasible sensitivity for measuring 2D/3D correlation spectra, but this time-consuming procedure and its associated expenses have restricted the application of ssNMR in lignocellulose analysis. Here, we present a method that relies on the sensitivity-enhancing technique Dynamic Nuclear Polarization (DNP) to eliminate the need for 13C-labeling. With a 26-fold sensitivity enhancement, a series of 2D 13C–13C correlation spectra were successfully collected using the unlabeled stems of wild-type Oryza sativa (rice). The atomic resolution allows us to observe a large number of intramolecular cross peaks for fully revealing the polymorphic structure of cellulose and xylan. NMR relaxation and dipolar order parameters further suggest a sophisticated change of molecular motions in a ctl1 ctl2 double mutant: both cellulose and xylan have become more dynamic on the nanosecond and microsecond timescale, but the motional amplitudes are uniformly small for both polysaccharides. By skipping isotopic labeling, the DNP strategy demonstrated here is universally extendable to all lignocellulose materials. This time-efficient method has landed the technical foundation for understanding polysaccharide structure and cell wall assembly in a large variety of plant tissues and species.
TL;DR: The results in this study provided valuable information to illustrate the molecular mechanisms of coordinate relations between astaxanthin and fatty acid biosynthesis and salicylic acid might play a role in self-protection processes of cells, helping adaption of H. pluvialis to high light stress.
Abstract: The unicellular alga Haematococcus pluvialis has achieved considerable interests for its capacity to accumulate large amounts of triacylglycerol and astaxanthin under various environmental stresses. To our knowledge, studies focusing on transcriptome research of H. pluvialis under exogenous hormones together with physical stresses are rare. In the present study, the change patterns at transcriptome level were analyzed to distinguish the multiple defensive systems of astaxanthin and fatty acid metabolism against exogenous salicylic acid and high light (SAHL) stresses. Based on RNA-seq data, a total of 112,463 unigenes and 61,191 genes were annotated in six databases, including NR, KEGG, Swiss-Prot, PFAM, COG and GO. Analysis of differentially expressed genes (DEGs) in KEGG identified many transcripts that associated with the biosynthesis of primary and secondary metabolites, photosynthesis, and immune system responses. Furthermore, 705 unigenes predicted as putative transcription factors (TFs) were identified, and the most abundant TFs families were likely to be associated with the biosynthesis of astaxanthin and fatty acid in H. pluvialis upon exposure to SAHL stresses. Additionally, majority of the fifteen key genes involved in astaxanthin and fatty acid biosynthesis pathways presented the same expression pattern, resulting in increased accumulation of astaxanthin and fatty acids in single celled H. pluvialis, in which astaxanthin content increased from 0.56 ± 0.05 mg·L−1 at stage Control to 0.89 ± 0.12 mg·L−1 at stage SAHL_48. And positive correlations were observed among these studied genes by Pearson Correlation (PC) analysis, indicating the coordination between astaxanthin and fatty acid biosynthesis. In addition, protein–protein interaction (PPI) network analysis also demonstrated that this coordination might be at transcriptional level. The results in this study provided valuable information to illustrate the molecular mechanisms of coordinate relations between astaxanthin and fatty acid biosynthesis. And salicylic acid might play a role in self-protection processes of cells, helping adaption of H. pluvialis to high light stress.
TL;DR: In this article, the authors investigated lipid accumulation in a variety of oleaginous ascomycetous and basidiomycete strains grown in glucose and xylose and followed lipid formation kinetics of selected strains in wheat straw hydrolysate.
Abstract: Microbial oils, generated from lignocellulosic material, have great potential as renewable and sustainable alternatives to fossil-based fuels and chemicals. By unravelling the diversity of lipid accumulation physiology in different oleaginous yeasts grown on the various carbon sources present in lignocellulose hydrolysate (LH), new targets for optimisation of lipid accumulation can be identified. Monitoring lipid formation over time is essential for understanding lipid accumulation physiology. This study investigated lipid accumulation in a variety of oleaginous ascomycetous and basidiomycetous strains grown in glucose and xylose and followed lipid formation kinetics of selected strains in wheat straw hydrolysate (WSH). Twenty-nine oleaginous yeast strains were tested for their ability to utilise glucose and xylose, the main sugars present in WSH. Evaluation of sugar consumption and lipid accumulation revealed marked differences in xylose utilisation capacity between the yeast strains, even between those belonging to the same species. Five different promising strains, belonging to the species Lipomyces starkeyi, Rhodotorula glutinis, Rhodotorula babjevae and Rhodotorula toruloides, were grown on undiluted wheat straw hydrolysate and lipid accumulation was followed over time, using Fourier transform-infrared (FTIR) spectroscopy. All five strains were able to grow on undiluted WSH and to accumulate lipids, but to different extents and with different productivities. R. babjevae DVBPG 8058 was the best-performing strain, accumulating 64.8% of cell dry weight (CDW) as lipids. It reached a culture density of 28 g/L CDW in batch cultivation, resulting in a lipid content of 18.1 g/L and yield of 0.24 g lipids per g carbon source. This strain formed lipids from the major carbon sources in hydrolysate, glucose, acetate and xylose. R. glutinis CBS 2367 also consumed these carbon sources, but when assimilating xylose it consumed intracellular lipids simultaneously. Rhodotorula strains contained a higher proportion of polyunsaturated fatty acids than the two tested Lipomyces starkeyi strains. There is considerable metabolic diversity among oleaginous yeasts, even between closely related species and strains, especially when converting xylose to biomass and lipids. Monitoring the kinetics of lipid accumulation and identifying the molecular basis of this diversity are keys to selecting suitable strains for high lipid production from lignocellulose.